Connectivity of Amphiprion akallopisos (skunk anemonefish) in
the Western Indian Ocean using microsatellite markers.
Annelore Van Nieuwenhove, Filip Huyghe, Marc Kochzius
anvnieuw@vub.ac.be Marine Biology, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium
INTRODUCTION
• Popular species in the pet and aquarium hobbyist trade (Fig. 1)
• Local overexploitation of populations and consequent destruction of coral reefs
• Networks of marine protected areas (MPAs) are necessary
• Connectivity among populations through larval dispersal should be taken into
account when designing MPAs
• This study: determining connectivity in the Western Indian Ocean (WIO) by
using microsatellites as genetic markers
Figure 1: Amphiprion akallopisos (photo: M. Kochzius)
Research questions:
1.
Is a genetic break present between the populations of Madagascar and the African mainland?
2.
Is there gene flow between the east and west coast of Madagascar?
3.
Are the results comparable when using mitochondrial DNA, a generally slower evolving genetic marker?
MATERIAL AND METHODS
SAMPLING
•
•
•
GENETIC ANALYSIS
Samples of 124 individuals
Collected at 6 different sites in
SCUBA-diving (Fig. 2)
Finclips were preserved in 96 % ethanol
the
WIO
during
• DNA extraction
• Multiplex PCR with four different fluorescent labels
applied to two sets of eight microsatellite markers (table 1)
• Scoring with program Genemarker
• Further DNA analysis with the programmes Genalex, Arlequin
and Structure.
Table 1: Applied microsatellite loci
Western Indian Ocean
LOCUS
PRIMER SEQUENCE
REPEAT
LENGTH (bp)
A130
F: GCACTCAACACAAAGACCTTA
R: ACCCAAACAACATCCAGTC
CA
200-300
D103
F: GTTGGCTAATGGTGCTGTG
R: GATTCTGTGGTGGCATCAG
GATA
200-250
B6
F: TGTCTTCTCCCAAGTCAG
R: ACGAGGCTCAACATACCTG
CATC
100-200
Am1
F: ACAAAGCCTTCATGTGGGTC
R: CGCAAGTGTTGCCTCATAGA
TG
100-150
Am6
F: AGCAGAGAGGAAAGAAGGGC
R: CAAGTGCCTGGCAGAAGATT
TGT
200-300
Am7
F: TGTCGCTACGACAGACTGCT
R: GCATGAGTGATTGGACCCTA
ATG
80-100
Am9
F: TGCTGCACTCTGTCTATTTTGT
R: GTGACTGAAGGCAAGGCAAT
TTA
200-250
Am10
F: GGAAGCAGCAATAAAGACGC
R: AGAGACGCCTGATGGTGAGT
ACAG
250-300
Am17
F: GGCTGTCTGGGATGAGATGT
R: TGTTCTGCAGATGGACTGTTTT
AATA
150-200
44
F: TTGGAGCAGCGTACTTAGCT
R: AGATGTGTTTACGCACGCTT
GT
200-250
61
F: TGAACACATAAACGCTCACTCAC
R: AAGACAATGCCTCCACATATCTA
GT
300-350
10TCTA
F: GGGACGTATCTGTTGGAAATGAT
R: TTAAGGTACTGTGAGATGAGACT
TCTA
500-600
CF11
F: GCTGGTTACAACACCTTG
R: GTAATTGCTGCAAGACAG
CT/CA
150-250
120
F: TCGATGACATAACACGACGCAGT
R: GACGGCCTCGATCTGCAAGCTGA
GT
400-500
AC137
F: GGTTGTTTAGGCCATGTGGT
R: TTGAGACACACTGGCTCCT
AC
350-450
D114
F: TGTTCCAGCTCTGATATTTGAC
R: TTGGCAGTGTTTTATACCTGTC
GATA
200-250
Figure 2: Map showing six sampling sites. Number of samples collected between brackets. (Google Maps)
Acknowledgement
We would like to thank the foundations that made this work possible: Fonds Wetenschappelijk Onderzoek Vlaanderen
for funding the project ‘Connectivity of Coral Reefs and Mangroves in the Western Indian Ocean’ (COCOMA-WIO; grant
1501612N); Vlaamse Interuniversitaire Raad - Universitaire Ontwikkelingssamenwerking (VLIR-UOS), for a travel grant
and scholarship to F.H, as well as travel grant to M.K.; Erasmus Mundus Programme CARIBU for a travel grant to M.K.;
H.A. Ratsimbazafy for providing samples from Madagascar; R.M. van der Ven (Vrije Universiteit Brussel), M. Sheikh and
M.S. Mohammed (State University of Zanzibar) and Jelvas M.Mwaura (Kenya Marine and Fisheries Research Institute) for
assistance during fieldwork; the competent authorities for permits.
The results of this study will contribute to the conservation and
management of coral reefs and its inhabitants.